GAL3ST1对肝细胞癌肿瘤进展的影响及机制研究

Effect and mechanism of GAL3ST1 on tumor progression of hepatocellular carcinoma

目的探究半乳糖-3-O-磺酰基转移酶1(GAL3ST1)对肝细胞癌(HCC)进展的影响及机制。方法以高转移人肝癌细胞株(MHCC97H)为研究对象,分别转染MHCC97H-GAL3ST1 OE上调GAL3ST1表达(GAL3ST1 OE组)、MHCC97H-GAL3ST1 KD下调GAL3ST1表达(GAL3ST1 KD组),另设不作任何处理的Ctrl组。采用MTT法检测各组细胞的增殖情况;流式细胞术检测各组细胞周期和凋亡情况;Western blotting检测鞘脂代谢通路蛋白的表达;观察小鼠皮下移植瘤的生长情况;实时荧光定量PCR(qPCR)检测各组UGT8和GALC mRNA的表达。结果培养24、48和72 h后,GAL3ST1 OE组细胞存活率分别为(127.27±2.47)%、(152.45±2.41)%和(174.62±0.78)%,均高于Ctrl组的(100±0.0)%、(125.86±1.84)%和(148.17±1.94)%,差异有统计学意义(P<0.05)。GAL3ST1 OE组S期细胞比例为(14.95±0.95)%,低于Ctrl组的(30.27±0.24)%,差异有统计学意义(P<0.05)。GAL3ST1 OE组早期及晚期细胞凋亡率分别(0.66±0.06)%和(1.51±0.15)%,低于Ctrl组的(2.85±0.21)%和(2.05±0.05)%,差异有统计学意义(P<0.05)。与Ctrl组比较,GAL3ST1 OE组Bax降低和Bcl-2、GALC和UGT8蛋白表达增加(P<0.05);GAL3ST1表达上调促进体内肝癌生长,促进Ki-67、PCNA、UGT8和GALC mRNA表达。结论GAL3ST1可能通过参与鞘脂代谢通路调控HCC进展。

Objective To investigate the effect and mechanism of galactose-3-O-sulfonyl transferase 1(GAL3ST1)on the progression of hepatocellular carcinoma(HCC).Methods The expression of GAL3ST1 was up-regulated by MHCC97h-GAL3ST1 OE and down-regulated by MHCC97h-GAL3ST1 KD,respectively.MTT was used to detect the proliferation of cells in each group.Cell cycle and apoptosis were detected by flow cytometry.Western Blotting was used to detect the expression of sphingolipid metabolic pathway proteins.The growth of subcutaneous transplanted tumor in mice was observed.The expression of UGT8 and GALC mRNA in each group was detected by real-time fluorescence quantitative PCR(qPCR).Results After 24,48 and 72 h culture,the survival rate of GAL3ST1 OE group was(127.27±2.47)%,(152.45±2.41)%and(174.62±0.78)%,which were higher than Ctrl group(100±0.0)%,(125.86±1.84)%and(148.17±1.94)%,the difference was statistically significant(P<0.05).The proportion of S-phase cells in GAL3ST1 OE group was(14.95±0.95)%,lower than that in Ctrl group(30.27±0.24)%,and the difference was statistically significant(P<0.05).The early and late apoptosis rates of GAL3ST1 OE group were(0.66±0.06)%and(1.51±0.15)%,respectively,which were lower than those of Ctrl group(2.85±0.21)%and(2.05±0.05)%,with statistical significance(P<0.05).Compared with Ctrl group,protein expression of Bax was decreased and protein expression of Bcl-2,GALC and UGT8 was increased in GAL3ST1 OE group(P<0.05).The up-regulated expression of GAL3ST1 promoted the growth of liver cancer in vivo,and promoted the mRNA expressions of Ki-67,PCNA,UGT8 and GALC.Conclusion GAL3ST1 may regulate the development of HCC by participating in sphingolipid metabolism pathway.